Ergovaline is produced by the endophyte Neotyphodium coenophialum (Morgan-Jones and Gams) in tall fescue (Schedonorus arundinacea (Schreb.) Dumort. = Festuca arundinacea Schreb.) and is blamed for a multitude of costly livestock disorders. Testing of pastures is common in both research and on farm situations. Since ergovaline is known to be unstable and affected by many variables, the objective of this study was to determine the effect of sample handling and storage on the stability of this compound. Homogeneous milled tall fescue sub-samples were analyzed for ergovaline concentration using HPLC after a range of sample handling procedures or storage. Ergovaline was unstable in milled material after 24 hours in storage, regardless of temperature. The decrease in ergovaline after 24 hours ranged from 17 to 60%. These results show that tall fescue sample handling and storage have a significant effect on ergovaline concentrations. In conclusion, accurate laboratory analysis of ergovaline content may require that samples be transported immediately to the laboratory on ice for immediate analysis. Most laboratories are not equipped for same day analysis, therefore researchers and producers should acknowledge that laboratory ergovaline results may be lower than the actual content in the field.

Tall fescue is a widely used forage grass in the eastern USA and can form a symbiosis with a fungal endophyte, which can be beneficial for the plant but can cause livestock health issues. Little is known regarding the symbiotic response to predicted climate change. To address this knowledge gap, I analyzed tall fescue variety trial data collected throughout the U.S., exploring relationships between climate variables and yield for two different fescue cultivars that were either endophyte-free or infected. This study showed no endophyte or cultivar effect on fescue yield, but identified temperature, precipitation and location as significant predictors of yield, suggesting that local conditions were more important than endophyte presence or fescue genotype for this dataset. Using a field experiment located in central Kentucky, I quantified the ecophysiological responses of four tall fescue genotypes to endophyte presence, elevated temperature and increased growing season precipitation. In this study, tall fescue genotype was as important as endophyte presence in determining ecophysiological responses to climate change treatments. My thesis illustrates that tall fescue response to climate change will depend on host genetics, the presence and genetics of the fungal endophyte symbiont, and the specific changes to the environment experienced at a site.

Perennial ryegrass (PRG, Lolium perenne) is a hardy cool-season grass that is infected with the endophytic fungus Neotyphodium lolii, which enables the plant to be insect repellant and drought resistant, lowering the use of insecticides and fertilizers. However, this fungus produces the compound lolitrem B (LB, m/z 686.4) which causes the tremorgenic neurotoxicity syndrome 'ryegrass staggers' in livestock consuming forage which contains <2000 ppb LB. Ergovaline (EV, m/z 534) is a vasoconstrictor normally associated with tall fescue (Festuca arudinacea), but has also been found in endophyte-infected PRG. Past research has shown a strong linear correlation between levels of LB and EV in PRG. The purpose of this study was to examine the linear relationship between EV and LB in feces and determine common metabolites. To accomplish this, four groups of steers (n=6/group) consumed endophyte- infected PRG over 70 days consumed the following averages of LB and EV: group I 2254ppb LB/633 ppb EV; group II 1554ppb LB/ 373ppb EV, group III 1011ppb LB/259ppb EV, and group IV 246ppb LB/<100ppb EV. Group I in week 4 was inadvertently given a washout period at which time the steers consumed the amount of LB and EV given to group IV (control). Both feed and feces samples were extracted using difference solid phase extraction methods and quantified by HPLC-fluorescence for LB and EV. Concentrations of EV and LB obtained through HPLC-fluorescence in both PRG and feces showed a linear relationship. Additional screening for metabolites was conducted LC-MS/MS and showed possible oxidation and reduction metabolites for both toxins. / Graduation date: 2013

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